Animal Imaging: CF Gene Therapy Departments of Radiology, Biomedical Engineering, and General medical Science Case Western Reserve University Cleveland, OH 44106 Zhenghong Lee, Ph.D.
Jan 12, 2016
Animal Imaging: CF Gene Therapy
Departments of Radiology, Biomedical Engineering, and General medical
Science
Case Western Reserve University
Cleveland, OH 44106
Zhenghong Lee, Ph.D.
Acknowledgement
CF Center (CWRU)
AG Ziady, Ph.D.PB Davis, M.D., Ph.D.
JLab (DOE)
S. Majewski, Ph.D.D. Weisenberger, Ph.D.
Radiology/BME (CWRU)
GR WojtkiewiczJJ DerakshanPF MartoneM Kotlarchyk
Univ. of Virginia
Dongfeng Pan, Ph.D.UCLA
H. Herschman, Ph.D.Q. Liang, Ph.D.
Molecular/Small Animal Imaging
Planar Multimodal Imaging System
Small animal scanner,developed by JLab at
CWRU/UHC, combining x-ray and scintigraphy
CF Gene Therapy Measurement
Molecular conjugate-DNA complex construction
+ + + +
+++
Plasmid DNA
Poly-L-lysine (~256 K)/LigandConjugate
DNA-Carrier Complex
aCl
Ethidium azide FITC
25 nm
C105Y
CF Studies: Gene Therapy
CF Studies: First Imaging
-IMAGER
Image Autoradiograph
CF Studies: Separate Imaging
Small animal imager with a highsensitivity copper collimator
Control with NaI Targeted delivery
CF Study: Validation
Reporter Gene
Suicide gene: HSV1-tk
HSV1-tk (gene) HSV1-TK (enzyme)
Ganciclovir (GCV) GCV-PO4
(cellular enzymes)
di/tri-PO4
Reporter gene: HSV1-tk Reporter gene probe: GCV
Reporter Gene for Imaging (Imagene)
Reporter gene: HSV1-tk
HSV1-tk (gene) HSV1-TK (enzyme)
[125I]FIAU FIAU-PO4
(cellular enzymes)
di/tri-PO4
Radiolabeled FIAU or other probes trappedto image reporter gene expression in vivo
TdR Analogues as Probe for HSV1-tk Expression
2’-fluoro-2’-deoxy-1--arabinofuranosyl-5-[I-125]-iodouracil (FIAU)
CF Studies: Multimodal Imaging
Dual modality imaging
CF Studies: Multimodal Imaging
Breathing mouse during imaging
125I-FIAU Injection to Image Stable HSV1-tk Complexes
Artifacts De-halogenation
HPLC Assay: [125I]-FIAU -trace
0.0 19.028 59.0
X-ray detectoruniformitycorrection
Blockingthyroid uptake
Imaging with Corrections
125I-FIAU Injection to Image Stable HSV1-tk Complexes
[I-125]-FIAU in Mouse Lungs
0
5
10
15
20
25
0.00 10.00 20.00 30.00 40.00 50.00 60.00 70.00 80.00 90.00
Time (post injection, hrs)
% o
f In
ject
ed D
ose
c1-both
c1-none
c4-both
c4-none
c5-right
c5-left
Furture Plans
Multimodality Imaging for CF Gene TherapyBioluminescent Imaging of Luc
Cancer Imaging Projects Peptide Ligand (none-RGD) for angiogenesis
Imaging Inflammatory Response Fab targeting neutrophils
Acknowledgement
Whitaker Foundation Imaging grant (RG-00-0305)
Cystic Fibrosis Foundation RDP grant (R477)
Department of Energy/JLab Subcontract (DE/SURA-04-Q011)
CF Studies: uPET Imaging (UCLA)
tk + luc (compacted) saline tk (compated)
CF Studies: uPET Imaging (UCLA)
microPET-measured sr39tk Expression in the Lung of Mice
0
0.5
1
1.5
2
0 1 2 3 4 5 6
Days (gene delivery at Day 0)
FH
BG
%ID
/g t
issu
e
tk(naked)
tk+luc(comp)
saline
tk(comp)
Two Concepts
Imaging Modalities
Anatomic Physiologic Metabolic Molecular
Nuclear (PET/SPECT)
CT
US
Optical Imaging
MR (MRI, MRS)
PET/SPECT Radio-nuclides